Muscella Antonella, Vetrugno Carla, Cossa Luca Giulio, Antonaci Giovanna, De Nuccio Francesco, De Pascali Sandra Angelica, Fanizzi Francesco Paolo, Marsigliante Santo
Laboratory of Cell Pathology, Department of Biological and Environmental Sciences and Technologies (Di.S.Te.B.A.), University of Salento, Lecce, Italy.
Laboratory of Cell Physiology Di.S.Te.B.A., University of Salento, Lecce, Italy.
PLoS One. 2016 Nov 2;11(11):e0165154. doi: 10.1371/journal.pone.0165154. eCollection 2016.
Malignant pleural mesothelioma (MPM) is an aggressive malignancy highly resistant to chemotherapy. There is an urgent need for effective therapy inasmuch as resistance, intrinsic and acquired, to conventional therapies is common. Among Pt(II) antitumor drugs, [Pt(O,O'-acac)(γ-acac)(DMS)] (Ptac2S) has recently attracted considerable attention due to its strong in vitro and in vivo antiproliferative activity and reduced toxicity. The purpose of this study was to examine the efficacy of Ptac2S treatment in MPM. We employed the ZL55 human mesothelioma cell line in vitro and in a murine xenograft model in vivo, to test the antitumor activity of Ptac2S. Cytotoxicity assays and Western blottings of different apoptosis and survival proteins were thus performed. Ptac2S increases MPM cell death in vitro and in vivo compared with cisplatin. Ptac2S was more efficacious than cisplatin also in inducing apoptosis characterized by: (a) mitochondria depolarization, (b) increase of bax expression and its cytosol-to-mitochondria translocation and decrease of Bcl-2 expression, (c) activation of caspase-7 and -9. Ptac2S activated full-length PKC-δ and generated a PKC-δ fragment. Full-length PKC-δ translocated to the nucleus and membrane, whilst PKC-δ fragment concentrated to mitochondria. Ptac2S was also responsible for the PKC-ε activation that provoked phosphorylation of p38. Both PKC-δ and PKC-ε inhibition (by PKC-siRNA) reduced the apoptotic death of ZL55 cells. Altogether, our results confirm that Ptac2S is a promising therapeutic agent for malignant mesothelioma, providing a solid starting point for its validation as a suitable candidate for further pharmacological testing.
恶性胸膜间皮瘤(MPM)是一种侵袭性恶性肿瘤,对化疗具有高度抗性。由于对传统疗法的固有和获得性抗性很常见,因此迫切需要有效的治疗方法。在铂(II)类抗肿瘤药物中,[Pt(O,O'-acac)(γ-acac)(DMS)](Ptac2S)最近因其强大的体外和体内抗增殖活性以及降低的毒性而备受关注。本研究的目的是检验Ptac2S治疗MPM的疗效。我们在体外使用ZL55人胸膜间皮瘤细胞系,并在体内使用小鼠异种移植模型,以测试Ptac2S的抗肿瘤活性。因此进行了细胞毒性测定以及不同凋亡和存活蛋白的蛋白质印迹分析。与顺铂相比,Ptac2S在体外和体内均增加了MPM细胞死亡。Ptac2S在诱导凋亡方面也比顺铂更有效,其特征为:(a)线粒体去极化,(b)bax表达增加及其从细胞质向线粒体的转位以及Bcl-2表达降低,(c)半胱天冬酶-7和-9的激活。Ptac2S激活全长PKC-δ并产生PKC-δ片段。全长PKC-δ转位至细胞核和细胞膜,而PKC-δ片段集中在线粒体。Ptac2S还导致PKC-ε激活,从而引起p38的磷酸化。PKC-δ和PKC-ε的抑制(通过PKC-siRNA)均降低了ZL55细胞的凋亡死亡。总之,我们的结果证实Ptac2S是一种有前途的恶性间皮瘤治疗药物,为其作为进一步药理测试的合适候选药物的验证提供了坚实的起点。